A recombinant lipoprotein antigen against Lyme disease expressed in E-coli:: Fermentor operating strategies for improved yield

被引：33

作者：

Madurawe, RD

Chase, TE

Tsao, EI

Bentley, WE ^{[1
]}

机构：

[1] Univ Maryland, Engn Res Ctr, Bioproc Scale Up Facil, College Pk, MD 20742 USA

[2] Univ Maryland, Dept Chem Engn, Rockville, MD 20850 USA

[3] Univ Maryland, Ctr Agr Biotechnol, Rockville, MD 20850 USA

[4] Medimmune Inc, Gaithersburg, MD 20878 USA

来源：

BIOTECHNOLOGY PROGRESS | 2000年 / 16卷 / 04期

关键词：

D O I：

10.1021/bp0000555

中图分类号：

Q81 [生物工程学（生物技术）]; Q93 [微生物学];

学科分类号：

071005 ; 0836 ; 090102 ; 100705 ;

摘要：

Decorin-binding lipoprotein, lpp-DBP, a bacterial surface adhesin, shows promise as a vaccine against Lyme disease. It is expressed in recombinant E. coli as an undesirable 20.5 KDa apoprotein that is subsequently lipidated in vivo to the desired 22 KDa lpp-DBP form. This study defines fermentation conditions for maximizing Ipp-DBP yield Super broth medium, a low post-induction temperature (30 degrees C), and a glucose feed based on dissolved oxygen resulted in high Ipp-DBP yield and minimized apoprotein formation. Since cells lysed within 2-3 h after induction, the cell yield was maximized by growing cells to high cell density prior to induction. Compared to a glucose feed based on maintaining a constant fermenter glucose concentration (Glucose-Stat), feeding based on maintaining a constant dissolved oxygen level (DO-Stat) improved yields. Also, a dissolved oxygen level of 60% (air saturation) was best, as no product degradation was detected by Western blotting and SDS-PAGE. Acetic acid levels under both modes of glucose feed were sufficiently low, and no adverse growth effects were observed.

引用

页码：571 / 576

页数：6

共 19 条

[1]

ATLAS RM, 1993, HDB MICROBIOLOGICAL, P281

[2]

BERGMEYER HU, 1974, METHOD ENZYMAT AN, V1, P112

[3] MOLECULAR ANALYSIS OF LINEAR PLASMID-ENCODED MAJOR SURFACE-PROTEINS, OSPA AND OSPB, OF THE LYME-DISEASE SPIROCHETE BORRELIA-BURGDORFERI [J].